Subsequent analyses of 551 CTCs confirmed the genetically homogeneous cancer in three serial blood analyses.
Inversely, 94% of the mutations present in cfDNA or CTC were confirmed in tDNA (Fig. 5a, b right Venn diagrams).
When a lesion detected at CT colonography (CTC) is not confirmed by a high quality colonoscopy, ESGE/ESGAR recommend careful review of the CTC findings.
In the present study, VEGF expression in CTCs was further confirmed by quantitative real-time RT-PCR in immunomagnetically separated CTCs.
Among 70 cfDNA and 39 CTC samples of overt myeloma samples (newly diagnosed or relapsed), there were 76%, 41%, and 24% of cfDNA samples with ≥3%, 5%, and 10% tumor fractions, respectively (Fig. 1c, blue panel).
Successful DNA extraction was confirmed in all samples with a qPCR targeting the human 18S rRNA gene with forward primer 5'- CTC TTA GCT GAG TGT CCC GC -3', reverse primer 5'- CTT AAT CAT GGC CTC AGT TCC GA -3', and probe labelled with fluorescein (FAM) and quenched with TAMRA: 5'-FAM CCG AGC CGC CTG GAT ACC GCA GCT A TAMRA-3'.
Overall, an average of 96% of non-silent clonal mutations that were present in cfDNA was confirmed in CTC, while 84% of non-silent clonal mutations that were present in CTC was confirmed in cfDNA.
Prior to use, drug concentrations in all CTC-96 solutions were confirmed by HPLC against an independently prepared CTC-96 standard.
MM_2205 CTC sample used 0.7 power due to lower coverage.
30/31 (96.8%) samples had at least 1 CTC, 15/31 (48.4%) samples at least 10 CTCs, with CTC total counts ranging from 0.1 to 9.67 CTCs/mL of blood (Table 1, Fig. 2B).
We applied our integrated workflow to 107 cfDNA samples and 56 CTC samples with different stages of disease progression.
